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Efficacy of Aqueous and Cow-Urine Based Biopesticides Against Journal of Pharmacognosy and Phytochemistry 2019; 8(3): 28-31 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2019; 8(3): 28-31 Efficacy of aqueous and cow-urine based Received: 17-03-2019 Accepted: 19-04-2019 biopesticides against Phytophthora nicotianae var. parasitica causing buckeye rot of tomato Shridhar BP Department of Plant Pathology, Dr. YS Parmar University of Horticulture and Forestry, Neri, Shridhar BP, Monica Sharma and Amit Sharma Hamirpur, Himachal Pradesh, India Abstract Buckeye rot of tomato, caused by Phytophthora nicotianae var. parasitica, is a serious threat to the crop Monica Sharma production and has taken a heavy toll of the crop in India which affects mostly the fruits during both Department of Plant Pathology, spring and winter season crops. In the present investigation, aqueous and cow-urine based leaf extracts of Dr. YS Parmar University of Horticulture and Forestry, Neri, five plants viz., Lantana camara, Artemisia vulgaris, Murraya koenigii, Eucalyptus tereticornis and Hamirpur, Himachal Pradesh, Melia azaderach were evaluated for their efficacy against the test pathogen under in vitro conditions. Out India of five aqueous biopesticides, Eucalyptus tereticornis was found most effective with 51.48 per cent inhibition in mycelial growth of the pathogen followed by Artemisia vulgaris (48.52%), Lantana camara Amit Sharma (44.81%) and Murraya koenigii (42.96%). Aqueous extract of Melia azedarach was least effective with Department of Basic Sciences, 39.62 per cent mycelial growth inhibition. Among five cow-urine based biopesticides, Eucalyptus College of Horticulture and tereticornis was found most effective with 71.89 per cent inhibition in mycelial growth of the pathogen Forestry, Dr. YS Parmar followed by Artemisia vulgaris (68.74%), Lantana camara (63.58%) and Murraya koenigii (59.51%). University of Horticulture and Cow-urine based botanical extract of Melia azedarach was least effective with 55.41 per cent mycelial Forestry, Neri, Hamirpur, growth inhibition. Himachal Pradesh, India Keywords: Plant extracts, antifungal, eco-friendly, indigenous, organic Introduction Tomato (Solanum lycopersicum L.), a member of family Solanaceae, is prone to a number of diseases among which, buckeye rot of unripe tomato is of serious concern to small and marginal hill farmers as it sometimes results in complete failure when fruiting coincides with the onset of rains. High relative humidity coupled with warm weather conditions causes high incidence of the disease. In India, losses due to this disease range between 35 and 40 per cent which may rise with the prevalence and severity of disease up to 90 per cent depending upon the favourable weather conditions. Under high humidity and good rainfall conditions, the incidence of buckeye rot alone may go up to 90 per cent (Gupta and Thind, 2006) [7]. The synthetic chemical fungicides are being used successfully for the management of diseases, but indiscriminate use of these chemicals led to have development of fungicide resistance in pathogen population and more important environmental pollution, posing a potential risk to animal and human health such as undesirable side effects due to their carcinogenic properties (Alkhail, 2005; Lyon et al., 1995) [1, 11]. Considering the adverse and alarming effects of synthetic pesticides on environment and natural habitats and the promotion of environmentally sustainable and organic agriculture, fungicide alternatives such as the use of natural plant [19, 15] products is needed (Slusarenko et al., 2008; Rice et al., 1998) . Plants contain antimicrobial compounds which can be toxic to pathogens and these natural plant products can be used to control plant diseases. Contrary to the problems associated with the use of synthetic chemicals, botanical extracts are environmentally non pollutive, renewable, inexhaustible, indigenously available, easily accessible, largely non phytotoxic, systemic ephemeral, thus readily biodegradable, relatively cost effective and hence constituted as suitable plant [10] protecting agents in the strategy of disease management (Kuberan et al., 2012) . The objective of this study was to screen leaf extracts of botanicals for antimicrobial activity against the Phytophthora nicotianae var. parasitica, a pathogen of tomato causing buckeye rot disease with the aim to develop alternative control strategy to reduce dependency on synthetic Correspondence Monica Sharma fungicides. Department of Plant Pathology, Dr. YS Parmar University of Materials and Methods Horticulture and Forestry, Neri, Source of pathogen Hamirpur, Himachal Pradesh, The pathogen was isolated from the infected fruits of tomato showing the typical symptoms of India buckeye rot disease following the standard isolation methods and was used as inoculum. ~ 28 ~ Journal of Pharmacognosy and Phytochemistry The pathogen isolated was maintained on carrot-juice agar medium throughout the study at 25 ± 1°C in an incubator for 7 days. Where, Plant materials and extract preparation I = Per cent inhibition Freshly harvested (200 g) leaves of plants viz., Lantana C = Growth of test pathogen in absence of biopesticide (cm) camara, Artemisia vulgaris, Murraya koenigii, Eucalyptus T = Growth of test pathogen in presence of biopesticide (cm) tereticornis and Melia azedarach were taken and washed under tap water. Each sample was grinded in mixer and In vitro evaluation of cow urine based bio-pesticides blender by adding small quantity of sterilized distilled water. Cow-urine based leaf extracts of Lantana camara, Artemisia After grinding, distilled water was added to make the final vulgaris, Murraya koenigii, Eucalyptus tereticornis and Melia volume 200 ml. It was then homogenized in orbital shaker for azedarach were evaluated under in vitro conditions against half an hour to get 100 per cent (w/v) botanical extract of each the pathogen by following poisoned food technique (Falck, plant. The extracts were then filtered through double-layered 1907; Nene and Thapliyal 2011) [6, 13]. Desired concentrations muslin cloth. Sterilization of botanical extracts was done of cow-urine based botanical extracts, each at 2, 5 and 10 per through tyndallization in autoclave at pressure 5 psi for 30 cent was prepared in sterilized distilled water by mixing with minutes for three consecutive days. The extracts were kept in equal quantity of double strength sterilized carrot-juice agar refrigerator for further use. In case of preparation of cow- medium and poured aseptically in sterilized Petriplates. urine based botanical extract, same procedure was followed as Medium mixed with cow urine only at 2, 5 and 10 per cent mentioned above but distilled water was replaced by ten days concentration was preapared along with medium mixed with old cow-urine to make the final volume of 200 ml of the equal quantities of sterilized distilled water without any botanical extract to make 100 per cent (w/v) cow urine based treatment were taken as control. These plates were then botanical extract. inoculated with mycelial discs of 5 mm diameter taken from actively growing culture of P. nicotianae var. parasitica in In vitro evaluation of aqueous biopesticides the centre and incubated at 25±1 °C. Experiment was Botanical extracts of Lantana camara, Artemisia vulgaris, conducted in completely randomized design (CRD) and each Murraya koenigii, Eucalyptus tereticornis and Melia treatment was replicated thrice. Regular observations were azedarach were evaluated under in vitro conditions against made and finally colony diameter was measured after the the pathogen by following poisoned food technique (Falck, [6, 13] control plates were completely covered by the pathogen. Per 1907; Nene and Thapliyal 2011) . Desired concentrations cent inhibition of mycelia growth was calculated by using of botanical extracts, each at 5, 10 and 15 per cent was formula (Vincent, 1947) [20]. prepared in sterilized distilled water by mixing with equal quantity of double strength sterilized carrot-juice agar Results medium and poured aseptically in sterilized Petriplates. Efficacy of aqueous biopesticides Medium mixed with equal quantities of sterilized distilled The results show that all the aqueous extracts of botanicals water without any treatment was taken as control. These inhibited the mycelial growth of P. nicotianae var. parasitica plates were then inoculated with mycelia discs of 5 mm in comparison to control. Aqueous extracts of Eucalyptus diameter taken from actively growing culture of P. nicotianae tereticornis was the most effective and significantly superior var. parasitica in the centre and incubated at 25±1 °C in BOD amongst all the treatments with 51.48 per cent inhibition in incubator. Each treatment was replicated thrice in completely mycelial growth of the pathogen followed by Artemisia randomized design (CRD). Regular observations were made vulgaris, Lantana camara and Murraya koenigii with 48.52, and finally colony diameter was measured after the control 44.81 and 42.96 per cent inhibition, respectively. Aqueous plates were completely covered by the pathogen. extract of Melia azedarach was least effective with 39.62 per Per cent inhibition of mycelial growth was calculated by the cent mycelial growth inhibition. formula (Vincent, 1947) [20]: Table 1: In vitro effect of aqueous biopesticides on the mycelial growth of P. nicotianae var. parasitica Per cent mycelial growth inhibition at different concentrations (%) Treatment 5 10 15 Mean Lantana camara 31.11 (33.87) 38.89 (38.56) 64.44 (53.38) 44.81 (41.93)c
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